Description
Principle of the assay: The coated well immunoenzymatic assay for the quantitative measurement of NTRK2 utilizes a multiclonal anti-NTRK2 antibody and an NTRK2-HRP conjugate. The assay sample and buffer are incubated together with NTRK2-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the NTRK2 concentration since NTRK2 from samples and NTRK2-HRP conjugate compete for the anti-NTRK2 antibody binding site. Since the number of sites is limited, as more sites are occupied by NTRK2 from the sample, fewer sites are left to bind NTRK2-HRP conjugate. Standards of known NTRK2 concentrations are run concurrently with the samples being assayed and a standard curve is plotted relating the intensity of the color (Optical Density) to the concentration of NTRK2. The NTRK2 concentration in each sample is interpolated from this standard curve